Molecular Modeling Approaches for Determining Gene Function: application to a Putative Poly-A Binding Protein from Leishmania amazonensis (LaPABP)

The great expansion in the number of genome sequencing projects has revealed the importance of computational methods to speed up the characterization of unknown genes. These studies have been improved by the use of three dimensional information from the predicted proteins generated by molecular modeling techniques. In this work, we disclose the structure-function relationship of a gene product from Leishmania amazonensis by applying molecular modeling and bioinformatics techniques. The analyzed sequence encodes a 159 aminoacids polypeptide (estimated 18 kDa) and was denoted LaPABP for its high homology with poly-A binding proteins from trypanosomatids. The domain structure, clustering analysis and a three dimensional model of LaPABP, basically obtained by homology modeling on the structure of the human poly-A binding protein, are described. Based on the analysis of the electrostatic potential mapped on the model's surface and conservation of intramolecular contacts responsible for folding stabilization we hypothesize that this protein may have less avidity to RNA than it's L. major counterpart but still account for a significant functional activity in the parasite. The model obtained will help in the design of mutagenesis experiments aimed to elucidate the mechanism of gene expression in trypanosomatids and serve as a starting point for its exploration as a potential source of targets for a rational chemotherapy.

The pathogenesis of diabetic complications: the role of DNA injury and poly(ADP-ribose) polymerase activation in peroxynitrite-mediated cytotoxicity

Recent work has demonstrated that hyperglycemia-induced overproduction of superoxide by the mitochondrial electron-transport chain triggers several pathways of injury [(protein kinase C (PKC), hexosamine and polyol pathway fluxes, advanced glycation end product formation (AGE)] involved in the pathogenesis of diabetic complications by inhibiting glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity. Increased oxidative and nitrosative stress activates the nuclear enzyme, poly(ADP-ribose) polymerase-1 (PARP). PARP activation, on one hand, depletes its substrate, NAD+, slowing the rate of glycolysis, electron transport and ATP formation. On the other hand, PARP activation results in inhibition of GAPDH by poly-ADP-ribosylation. These processes result in acute endothelial dysfunction in diabetic blood vessels, which importantly contributes to the development of various diabetic complications. Accordingly, hyperglycemia-induced activation of PKC and AGE formation are prevented by inhibition of PARP activity. Furthermore, inhibition of PARP protects against diabetic cardiovascular dysfunction in rodent models of cardiomyopathy, nephropathy, neuropathy, and retinopathy. PARP activation is also present in microvasculature of human diabetic subjects. The present review focuses on the role of PARP in diabetic complications and emphasizes the therapeutic potential of PARP inhibition in the prevention or reversal of diabetic complications.

The effects of drugs, ions, and poly-l-lysine on the excretory system of Schistosoma mansoni

We have been able to label the excretory system of cercariae and all forms of schistosomula, immature and adult worms with the highly fluorescent dye resorufin. We have shown that the accumulation of the resorufin into the excretory tubules and collecting ducts of the male adult worm depends on the presence of extracellular calcium and phosphate ions. In the adult male worms, praziquantel (PZQ) prevents this accumulation in RPMI medium and disperses resorufin from tubules which have been prelabelled. Female worms and all other developmental stages are much less affected either by the presence of calcium and phosphate ions, or the disruption caused by PZQ. The male can inhibit the excretory system in paired female. Fluorescent PZQ localises in the posterior gut (intestine) region of the male adult worm, but not in the excretory system, except for the anionic carboxy fluorescein derivative of PZQ, which may be excreted by this route. All stages of the parasite can recover from damage by PZQ treatment in vitro. The excretory system is highly sensitive to damage to the surface membrane and may be involved in vesicle movement and damage repair processes. In vivo the adult parasite does not recover from PZQ treatment, but what is inhibiting recovery is unknown, but likely to be related to immune effector molecules.

Ammonium chloride as nitrogen source in sugarcane harvested without burning

The great difficulty of incorporation of N fertilizers into the "green sugarcane" system causes concern and since urea is the most commonly used source, there is the risk of loosing NH3 through volatilization. For this reason, a field experiment was undertaken (in a Hapludox Typic) with the objective of evaluating the agronomic efficiency of ammonium chloride on stubble of the second ratoon (SP89 1115), as well as its residual effect on the subsequent cycle (third ratoon). The experimental design was randomized blocks with four replications. Treatments consisted of three N rates (60, 120 and 180 kg ha-1 N) in the form of NH4Cl, in addition to a control treatment without the addition of N fertilizer. The ratoon cane of the second cutting was harvested in November 2006 and the treatments were applied in December 2006. The second ratoon was harvested mechanically in November 2007 and in December 2007, 450 kg ha-1 of the NPK mixture 20-05-19 was applied, providing 90, 22 and 86 kg ha-1 N, P2O5 and K2O, respectively, for the purpose of evaluating the effect of residual-N from the treatments implanted in December 2006. An increase in the rates of N-NH4Cl had a positive effect on the leaf concentrations of P, Mg and S. Stalk yield (MSS - Mg ha-1 of sugarcane stalks) and sugar (MSH - Mg ha-1 of sucrose) in the November 2006 harvest responded linearly to the increase of N doses in the form of NH4Cl. In relation to the effect of residual-N in the 2007/2008 harvest, it was observed, in general, that the concentrations of macronutrients in the sugarcane leaf +1 were within the range considered adequate in the state of São Paulo, Brazil. The residual-N of the NH4Cl doses resulted in a significant reduction in stalk (MSS) and sugar (MSH) production. It may be concluded that the NH4Cl source at a dose of 120 kg ha-1 N in ratoon fertilization of the second cutting was agronomically efficient, presenting, however, less efficiency of residual-N in the subsequent cycle.

Application of 1-methylcyclopropene, calcium chloride and calcium amino acid chelate on fresh-cut cantaloupe muskmelon

The objective of this work was to determine the effects of postharvest application of 1-methylcyclopropene (1-MCP) and two calcium salts, applied individually or combined, on firmness and visual quality of fresh-cut muskmelon stored in air, for 18 days. Two sets of fruits, one of them exposed to 1-MCP at 300 nL L-1, were cut into cubes, dipped in deionized water, or in 1% Ca solutions as CaCl2, or in calcium amino acid chelate (Ca-chelate), placed in clamshell containers, and stored in air at 5±1ºC and 90±5% RH, for 18 days. The assay was conducted using an entirely randomized design, with three replications, in a split plot array. Evaluation of visual appearance, color, flesh firmness, total soluble solids, titratable acidity, and pH was performed right after treatments, and every period of three days, up to eighteen days. Application of 1-MCP at 300 nL L-1, calcium chloride or Ca-chelate, or the combination 1-MCP and calcium, preserved initial freshness and reduced softening of the samples. Ca-chelate synergistically enhanced the effect of 1-MCP on firmness after nine days of storage, while calcium chloride improved firmness of the samples throughout storage. Ca-chelate may serve as an alternative for shelf life extension of cantaloupe fresh-cut muskmelon.

Potasssium chloride as a nutrient seed primer to enhance salt‑tolerance in maize

The objective of this work was to determine if KCl could be a useful nutrient primer for safe seed germination in maize crop under salt stress conditions. Seed priming was done using 50 mmol L‑1 of muriate of potash, and germination and seedling growth were evaluated after salt stress with NaCl up to 50 mmol L‑1. Another set of seeds was tested under the same salt stress conditions without priming. Under salinity stress, germination percentage, germination rate index, germination coefficient, and seedling vigor indexes were higher in primed seeds. In unprimed seeds, mean germination time increased, while the germination rate index and the fresh and dry matter mass decreased more sharply with salinity stress. The Na/K ratio was higher in unprimed seeds.

Stereoselective sodium borohydride reductions of cyclopentanones: influence of ceric chloride on the stereochemistry of reaction

In this paper we describe the reduction by NaBH4 of some cyclopentanones containing an oxygenated function at the side chain position beta to the carbonyl group, both in the presence and in the absence of CeCl3. Some suggestions for the rationalization of the results are discussed, considering the stereochemical course of the reactions.

Synthesis and NMR characterization of aliphatic-aromatic copolyesters by reaction of poly(ethylene terephthalate) post-consumer and poly(ethylene adipate)

An aliphatic-aromatic copolyester of poly(ethylene terephthalate), PET, and poly(ethylene adipate), PEA, PET-co-PEA, was synthesized by the high temperature melt reaction of post-consumer PET and PEA. As observed by NMR spectroscopy, the reaction yielded random copolyesters in a few minutes through ester-interchange reactions, even without added catalyst. The copolyesters obtained in the presence of a catalyst presented higher intrinsic viscosity than that obtained without the addition of catalyst, due to simultaneous polycondensation and ester-interchange reactions. The structure of the aliphatic-aromatic copolyesters obtained in different PET/PEA ratio is random as observed by NMR analysis.

Preparação e caracterização óptica de filmes de poli(estireno sulfonados) dopados com neodímio

In this work we describe the processing of poly(styrene sulphonate) films (PSS) doped with neodymium (Nd). Optical density measurements in the UV-Vis-NIR region show the typical bands observed for neodymium chloride (NdCl3) in solution. In the case of films, the intensity ratio between the peaks at 800 nm (4I9/2 -> 4F5/2 + ²H7/2) and 580 nm (4I9/2 -> 4G5/2 + ²G7/2) is equal to 0.83. Infrared spectra present an enhancement in the absorption region of aromatic rings. Site selective luminescence spectroscopy shows that the incorporation of Nd introduces a hipsochromic shift and a line shape definition in UV luminescence compared to PSS film, decreasing the interaction between aromatic groups. In addition, the film exhibits an intense radiative transition at 1061 nm (4F3/2->4I11/2), comparable to the one present in crystalline materials doped with Nd.

Effect of the structure of commercial poly(ethylene oxide-b-propylene oxide) demulsifier bases on the demulsification of water-in-crude oil emulsions: elucidation of the demulsification mechanism

Water-in-crude oil emulsions are formed during petroleum production and asphaltenes play an important role in their stabilization. Demulsifiers are added to destabilize such emulsions,however the demulsification mechanism is not completely known. In this paper, the performances of commercial poly(ethylene oxide-b-propylene oxide) demulsifiers were studied using synthetic water-in-oil emulsions and model-systems (asphaltenes in organic solvent). No change in the asphaltene aggregate size induced by the demulsifier was observed. The demulsification performance decreased as the asphaltene aggregate size increased, so it can be suggested that the demulsification mechanism is correlated to the voids between the aggregates adsorbed on the water droplets surface.

Preparation of PVP hydrogel nanoparticles using lecithin vesicles

Hydrogels micro, sub-micro and nanoparticles are of great interest for drug encapsulation and delivery or as embolotherapic agents. In this work it is described the preparation of nano and sub-microparticles of pre-formed, high molecular weight and monomer free poly(N-vinyl-2-pyrrolidone) encapsulated inside the core of lecithin vesicles. The hydrogel particles are formed with a very narrow diameter distribution, of about 800 nm, and a moderate swelling ratio, of approximately 10.

Development of a flotation-spectrophotometric method for determination of cetylpyridinium chloride in pharmaceutical products

A new simple and sensitive flotation-spectrophotometric method for the determination of cetylpyridinium chloride (CPC) is reported. The method is based on the formation of an ion- associate between CPC and Orange II (OR) which is floated in the interface of aqueous phase and n-hexane by vigorous shaking. The aqueous solution was discarded and the adsorbed ion associate on to the wall of a separating funnel was dissolved in a small volume of methanol solvent and its absorbance was measured at 480 nm. The apparent molar absorptivity (Ε) of the ion associate was determined to be 4.12 x 10(5) L mol-1 cm-1. The calibration graph was linear in the concentration range of 15-800 ng mL-1 of CPC with a correlation coefficient of 0.9988. The limit of detection (LOD) was 10.8 ng mL-1. The relative standard deviation (RSD) for determination of 100 and 800 ng mL-1 of CPC was 3.47 and 2.04% (n=7), respectively. The method was successfully applied to the determination of CPC in a commercial mouth washer product.

Poly(ethylene-co-methyl acrylate)/poly(caprolactone) triol blends for drug delivery systems: characterization and drug release

Poly(ethylene-co-methyl acrylate) (EMA) and poly (caprolactone) triol (PCL-T) blends, a biodegradable aliphatic polyester with low molecular weight and moderate water solubility containing diltiazem hydrochloride (DZ) were studied in terms of the thermal and morphological properties, and drug release mechanism. An increase in the PCL-T content in the EMA/PCL-T/DZ films decreased the degree of DZ crystallinity. Drug release from these films is temperature-dependent, and it is possible to modify the drug release rate by adjusting the EMA/PCL-T composition of the blends. The mechanism of drug release is governed by PCL-T melting and PCL-T leaching from EMA matrix.

Poly(o-methoxyaniline) modified electrode for detection of lithium ions

This paper reports the use of an electrode modified with poly(o-methoxyaniline) for detecting lithium ions. These ions are present in drugs used for treating bipolar disorder and that requires periodical monitoring of the concentration of lithium in blood serum. Poly(o-methoxyaniline) was obtained electrochemically by cyclic voltammetry on the surface of a gold electrode. The results showed that the electrode modified with a conducting polymer responded to lithium ions in the concentration range of 1 x 10-5 to 1 x 10-4 mol L-1 . The results also confirmed that the performance of the modified electrode was comparable to that of the standard method (atomic emission spectrophotometry).